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pCS2+hSpCas9
(Plasmid #51815)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 51815 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 51815-D50 50 μg of DNA in Tris buffer $495
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DNA 51815-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pCS2
  • Backbone size w/o insert (bp) 4071
  • Total vector size (bp) 8343
  • Modifications to backbone
    Cloned in hSpCas9 expression sequence
  • Vector type
    Mammalian Expression, CRISPR ; medaka, zebrafish

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    humanized S.pyogenes Cas9
  • Alt name
    SpCas9
  • Alt name
    hSpCas9
  • Insert Size (bp)
    4272
  • Promoter SP6
  • Tag / Fusion Protein
    • 3xFLAG (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site Xba I (not destroyed)
  • 5′ sequencing primer SP6
  • 3′ sequencing primer T3
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Dr. Zhang produced the hSpCas9 gene, which has been deposited to Addgene (Plasmid 42230: pX330-U6-Chimeric_BB-CBh-hSpCas9 | Cong et al. Science. 3 Jan 2013).

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid is modified from pX330-U6-Chimeric BB-CBh-hSpCas9 originated from Feng Zhang Lab.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCS2+hSpCas9 was a gift from Masato Kinoshita (Addgene plasmid # 51815 ; http://n2t.net/addgene:51815 ; RRID:Addgene_51815)

  • For your References section:

    Targeted mutagenesis using CRISPR/Cas system in medaka. Ansai S, Kinoshita M. Biol Open. 2014 Apr 11;3(5):362-71. doi: 10.1242/bio.20148177. 10.1242/bio.20148177 PubMed 24728957
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